Di Lello Paola, Benison Gregory C, Valafar Homayoun, Pitts Keith E, Summers Anne O, Legault Pascale, Omichinski James G
Department of Biochemistry & Molecular Biology, University of Georgia, Athens, Georgia 30602, USA.
Biochemistry. 2004 Jul 6;43(26):8322-32. doi: 10.1021/bi049669z.
Mercury resistant bacteria have developed a system of two enzymes (MerA and MerB), which allows them to efficiently detoxify both ionic and organomercurial compounds. The organomercurial lyase (MerB) catalyzes the protonolysis of the carbon-mercury bond resulting in the formation of ionic mercury and a reduced hydrocarbon. The ionic mercury [Hg(II)] is subsequently reduced to the less reactive elemental mercury [Hg(0)] by a specific mercuric reductase (MerA). To better understand MerB's unique enzymatic activity, we used nuclear magnetic resonance (NMR) spectroscopy to determine the structure of the free enzyme. MerB is characterized by a novel protein fold consisting of three noninteracting antiparallel beta-sheets surrounded by six alpha-helices. By comparing the NMR data of free MerB and the MerB/Hg/DTT complex, we identified a set of residues that likely define a Hg/DTT binding site. These residues cluster around two cysteines (C(96) and C(159)) that are crucial to MerB's catalytic activity. A detailed analysis of the structure revealed the presence of an extensive hydrophobic groove adjacent to this Hg/DTT binding site. This extensive hydrophobic groove has the potential to interact with the hydrocarbon moiety of a wide variety of substrates and may explain the broad substrate specificity of MerB.
抗汞细菌已形成一种由两种酶(MerA和MerB)组成的系统,这使它们能够有效地将离子型和有机汞化合物解毒。有机汞裂解酶(MerB)催化碳-汞键的质子解,生成离子汞和还原态烃。离子汞[Hg(II)]随后被一种特定的汞还原酶(MerA)还原为活性较低的元素汞[Hg(0)]。为了更好地理解MerB独特的酶活性,我们使用核磁共振(NMR)光谱法来确定游离酶的结构。MerB的特征是具有一种新颖的蛋白质折叠结构,由三个不相互作用的反平行β-折叠组成,周围环绕着六个α-螺旋。通过比较游离MerB和MerB/Hg/DTT复合物的NMR数据,我们确定了一组可能定义Hg/DTT结合位点的残基。这些残基聚集在对MerB催化活性至关重要的两个半胱氨酸(C(96)和C(159))周围。对该结构的详细分析揭示,在这个Hg/DTT结合位点附近存在一个广泛的疏水凹槽。这个广泛的疏水凹槽有可能与多种底物的烃基部分相互作用,这可能解释了MerB广泛的底物特异性。